Team:Ionis Paris/Protocol 11
Following steps will be performed under a class II PSM Prepare a 50% glycerol stock solution by mixing 5mL glycerol with 5mL distilled, sterile water. Mix 50µL of the overnight culture of bacteria of interest with 50µL 50% glycerol solution (25% glycerol final concentration) into a 2mL cryotube Pipet bacteria cells and put it into a 500mL sterile Erlenmeyer containing 100mL of sterile liquid LB medium. Cover with aluminum paper and place the Erlenmeyer in a rotative incubator at 37 Celsius degrees, 120rpm O/N.
Protocol 11 : Glycerol storage
Aim: Safely store cells at -80°C
Glycerol Freezing Solution (50%)
Mix well to make sure that you see one uniform solution, and there are no layers present.
Bacteria storage
NB : Snap top tubes are not recommended as they can open unexpectedly at -80°C
Label both the lid and the tube of the cryotube. Frozen tubes are hard to write on and samples stored for long periods at -80°C can lose labels stuck to tube.
Store at -80°C
Bacteria recovery
